Th17/Treg balance and macrophage polarization ratio in lower extremity arteriosclerosis obliterans

Objective:To explore the balance of peripheral blood T helper 17 cells/regulatory T cell(Th17/Treg)ratio and the polarization ratio of M1 and M2 macrophages in lower extremity arteriosclerosis obliterans(ASO).Methods:A rat model of lower extremity ASO was established,and blood samples from patients with lower extremity ASO before and after surgery were obtained.ELISA was used to detect interleukin 6(IL-6),IL-10,and IL-17.Real-time RCR and Western blot analyses were used to detect Foxp3,IL-6,IL-10,and IL-17 expression.Moreover,flow cytometry was applied to detect the Th17/Treg ratio and M1/M2 ratio.Results:Compared with the control group,the iliac artery wall of ASO rats showed significant hyperplasia,and the concentrations of cholesterol and triglyceride were significantly increased(P<0.01),indicating the successful establishment of ASO.Moreover,the levels of IL-6 and IL-17 in ASO rats were pronouncedly increased(P<0.05),while the IL-10 level was significantly decreased(P<0.05).In addition to increased IL-6 and IL-17 levels,the mRNA and protein levels of Foxp3 and IL-10 in ASO rats were significantly decreased compared with the control group.The Th17/Treg and M1/M2 ratios in the ASO group were markedly increased(P<0.05).These alternations were also observed in ASO patients.After endovascular surgery(such as percutaneous transluminal angioplasty and arterial stenting),all these changes were significantly improved(P<0.05).Conclusions:The Th17/Treg and M1/M2 ratios were significantly increased in ASO,and surgery can effectively improve the balance of Th17/Treg,and reduce the ratio of M1/M2,and the expression of inflammatory factors.

Neutrophil peptide 1 accelerates the clearance of degenerative axons during Wallerian degeneration by activating macrophages after peripheral nerve crush injury

Macrophages play an important role in peripheral nerve regeneration,but the specific mechanism of regeneration is still unclear.Our preliminary findings indicated that neutrophil peptide 1 is an innate immune peptide closely involved in peripheral nerve regeneration.However,the mechanism by which neutrophil peptide 1 enhances nerve regeneration remains unclear.This study was designed to investigate the relationship between neutrophil peptide 1 and macrophages in vivo and in vitro in peripheral nerve crush injury.The functions of RAW 264.7 cells we re elucidated by Cell Counting Kit-8 assay,flow cytometry,migration assays,phagocytosis assays,immunohistochemistry and enzyme-linked immunosorbent assay.Axonal debris phagocytosis was observed using the CUBIC(Clear,Unobstructed Brain/Body Imaging Cocktails and Computational analysis)optical clearing technique during Wallerian degeneration.Macrophage inflammatory factor expression in different polarization states was detected using a protein chip.The results showed that neutrophil peptide 1 promoted the prolife ration,migration and phagocytosis of macrophages,and CD206 expression on the surfa ce of macrophages,indicating M2 polarization.The axonal debris clearance rate during Wallerian degeneration was enhanced after neutrophil peptide 1 intervention.Neutrophil peptide 1 also downregulated inflammatory factors interleukin-1α,-6,-12,and tumor necrosis factor-αin invo and in vitro.Thus,the results suggest that neutrophil peptide 1 activates macrophages and accelerates Wallerian degeneration,which may be one mechanism by which neutrophil peptide 1 enhances peripheral nerve regeneration.

Effects of interleukin-10 treated macrophages on bone marrow mesenchymal stem cells via signal transducer and activator of transcription 3 pathway

BACKGROUND Alveolar bone defects caused by inflammation are an urgent issue in oral implant surgery that must be solved.Regulating the various phenotypes of macrophages to enhance the inflammatory environment can significantly affect the progression of diseases and tissue engineering repair process.AIM To assess the influence of interleukin-10(IL-10)on the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)following their interaction with macrophages in an inflammatory environment.METHODS IL-10 modulates the differentiation of peritoneal macrophages in Wistar rats in an inflammatory environment.In this study,we investigated its impact on the proliferation,migration,and osteogenesis of BMSCs.The expression levels of signal transducer and activator of transcription 3(STAT3)and its activated form,phos-phorylated-STAT3,were examined in IL-10-stimulated macrophages.Subsequently,a specific STAT3 signaling inhibitor was used to impede STAT3 signal activation to further investigate the role of STAT3 signaling.RESULTS IL-10-stimulated macrophages underwent polarization to the M2 type through substitution,and these M2 macrophages actively facilitated the osteogenic differentiation of BMSCs.Mechanistically,STAT3 signaling plays a crucial role in the process by which IL-10 influences macrophages.Specifically,IL-10 stimulated the activation of the STAT3 signaling pathway and reduced the macrophage inflammatory response,as evidenced by its diminished impact on the osteogenic differentiation of BMSCs.CONCLUSION Stimulating macrophages with IL-10 proved effective in improving the inflammatory environment and promoting the osteogenic differentiation of BMSCs.The IL-10/STAT3 signaling pathway has emerged as a key regulator in the macrophage-mediated control of BMSCs’osteogenic differentiation.

Toxoplasma ROP16Ⅰ/Ⅲ ameliorated inflammatory bowel diseases via inducing M2 phenotype of macrophages

BACKGROUND Inflammatory bowel disease(IBD)is characterized by chronic and non-specific inflammation of the intestinal mucosa and mainly includes ulcerative colitis and Crohn's disease.AIM To explore the beneficial effect of ToxoROP16I/III-induced M2 phynotype macrophages in homeostasis of IBDs through downregulation of M1 inflammatory cells.METHODS RAW264.7 macrophages stimulated by lipopolysaccharide(LPS)(M1 cells)were co-cultured with Caco-2 cells as an inflammatory model of IBD in vitro.The expression of ToxoROP16I/III was observed in RAW264.7 macrophages that were transfected with pEGFP-rop16I/III.The phenotypes of M2 and M1 macrophage cells were assessed by quantitative real-time reverse transcriptase polymerase chain reaction and the expression of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,transforming growth factor(TGF)-β1,IL-10,inducible nitric oxide synthase(iNOS),and arginase-1(Arg-1)was detected.The expression of iNOS,Arg-1,signal transducer and activator of transcription 3(Stat3),p-Stat3,Stat6,p-Stat6,programmed death ligand-2(PD-L2),caspase-3,-8,and-9 was analyzed by Western blotting,and Griess assays were performed to detect nitric oxide(NO).TNF-α,IL-1β,IL-6,TGF-β1,and IL-10 expression in the supernatants was detected by enzyme-linked immunosorbent assay,and Caco-2 cell apoptosis was co-culture system.RESULTS M1 cells exhibited significantly increased production of iNOS,NO,TNF-α,IL-1β,and IL-6,while ToxoROP16I/III induced macrophage bias to M2 cells in vitro,showing increased expression of Arg-1,IL-10 and TGF-β1 and elevated production of p-Stat3 and p-Stat6.The mixed M1 and M2 cell culture induced by ToxoROP16I/III exhibited decreased production of NO and iNOS and upregulated expression of Arg-1 and PD-L2.Accordingly,Caco-2 cells became apoptotic,and apoptosis-associated proteins such as caspase-3,-8 and-9 were dampened during co-culture of M1 and M2 cells.Flow cytometry analysis showed that co-culture of M1 cells with Caco-2 cells facilitated the apoptosis of Caco-2 cells,but co-culture of M1 and M2 cells alleviated Caco-2 cell apoptosis.CONCLUSION ToxoROP16I/III-induced M2 macrophages inhibited apoptosis of Caco-2 cells caused by M1 macrophages.This finding may help gain a better understanding of the underlying mechanism and represent a promising therapeutic strategy for IBDs.

The Expression of Interleukin-17, Interferon-gamma, and Macrophage Inflammatory Protein-3 Alpha mRNA in Patients with Psoriasis Vulgaris

Summary: To investigate the role of Interleukin-17 (IL-17), Interferon-gamma (IFN-γ), and macrophage inflammatory protein-3 alpha (MIP-3α) in the pathogenesis of psoriasis, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to semi-quantitatively analyze the mRNA expression of IL-17, IFN-γ, and MIP-3α in 31 psoriatic lesions and 16 normal skin tissues. The results showed that the mRNA of the three cytokines was present in all specimens. And the expression level of IL-17 mRNA in skin lesions was 1.1416±0.0591, which was significantly higher than that in normal controls (0.8788±0.0344, P<0.001). The expression levels of IFN-γ mRNA were 1.1142±0.0561 and 0.9050±0.0263, respectively, with significant difference(P<0.001). And the expression levels of MIP-3α mRNA in psoriatic lesions was 1.1397±0.0521, which was markedly higher than that in normal controls (0.8681±0.0308, P<0.001). These findings indicate that up-regulated expression of IL-17, IFN-γ, and MIP-3α might be involved in the pathogenesis of psoriasis.

Suppression of Breast Cancer Proliferation and Induction of Apoptosis via AKT and ERK1/2 Signal Transduction Pathways by Synthetic Polypeptide Derived from Viral Macrophage Inflammatory Protein Ⅱ

SDF-1α,a ligand for the chemokine receptor CXCR4,is well known for mediating the migration of breast cancer cells.In a previous study we demonstrated that a synthetic 21-mer peptide antagonist of CXCR4（NT21MP） derived from the viral macrophage inflammatory protein Ⅱ could antagonize tumor growth in vivo by inhibiting cellular proliferation and inducing apoptosis in breast cancer cells.However,the role of SDF-1α in the signaling pathways underlying the proliferation of human breast cancer cells and associated signaling pathways and inhibiting signal pathways of NT21MP remained unclear.The present study investigated the mechanism of NT21MP on anti-tumor in breast cancer in vitro.The effect of NT21MP on the viability of cells was determined by the MTT assay.Annexin V-FITC and PI staining was performed to detect early stage apoptosisin SKBR3 cells treated with SDF-1α and AMD3100 or NT21MP.Western blotting techniques were used to assay the composition of phosphoproteomics and total proteins present in the SKBR3 breast cancer cells.RT-PCR and Western blotting technique were used to detect the effect of NT21MP and AMD3100 on Bcl-2 and Bax expression.The results indicated that SDF-1α prevented apoptosis and promoted the proliferation of SKBR3 human breast cancer cells.As compared with untreated SKBR3 cells,Treatment with SDF-1α significantly increased cell viability,and NT21MP abolished the protective effects of SDF-1α dose-dependently（P0.05）.There was a significant decrease in the percentage of apoptotic cells after SDF-1α treatment as compared with control group（2.7%±0.2% vs.5.7%±0.4%,P0.05）.But pretreatment of SKBR3 cells with NT21MP significantly attenuated the antiapoptotic effects of SDF-1α as compared with SKBR3 cells without NT21MP pretreatment.The proliferative and anti-apoptotic effects of SDF-1α in SKBR3 cells were associated with an increase in AKT and ERK1/2 phosphorylation as well as a decrease in Bax expression and an increase in Bcl-2 expression.These changes in intracellular processes were blocked by NT21MP in a dose-dependent manner（P0.05）.In conclusion,NT21MP efficiently inhibits SDF-1α-induced proliferation and antiapoptosis in SKBR3 cells by reducing the levels of phosphorylated AKT and ERK1/2,as well as decreasing the ratio of expression of Bcl-2 relative to Bax.

Macrophage inflammatory protein-2 as mediator of inflammation in acute liver injury

Macrophage inflammatory protein(MIP)-2 is one of the CXC chemokines and is also known as chemokine CXC ligand(CXCL2). MIP-2 affects neutrophil recruitment and activation through the p38 mitogen-activatedprotein-kinase-dependent signaling pathway, by binding to its specific receptors, CXCR1 and CXCR2. MIP-2 is produced by a variety of cell types, such as macrophages, monocytes, epithelial cells, and hepatocytes, in response to infection or injury. In liver injury, activated Kupffer cells are known as the major source of MIP-2. MIP-2-recruited and activated neutrophils can accelerate liver inflammation by releasing various inflammatory mediators. Here, we give a brief introduction to the basic molecular and cellular sources of MIP-2, and focus on its physiological and pathological functions in acute liver injury induced by concanavalin A, lipopolysaccharides, irradiation, ischemia/reperfusion, alcohol, and hypoxia, and hepatectomy-induced liver regeneration and tumor colorectal metastasis. Further understanding of the regulatory mechanisms of MIP-2 secretion and activation may be helpful to develop MIP-2-targeted therapeutic strategies to prevent liver inflammation.

Macrophage Inflammatory Protein-1 Beta (MIP-1<i>β</i>) and Platelet Indices as Predictors of Spontaneous Bacterial Peritonitis<br>—MIP, MPV and PDW in SBP

Background/Aims: The objective of this study is to measure macrophage inflammatory protein one beta (MIP-1β), mean platelet volume (MPV) and platelet distribution width (PDW) to evaluate their usefulness in the diagnosis of spontaneous bacterial peritonitis (SBP) in cirrhotic patients. Materials and Methods: This study comprised 41 cirrhotic patients with ascites. MPV, PDW and MIP-1β were measured in serum and ascitic fluid. Results: A significant increase MPV, PDW, C-reactive Protein (CRP) and white blood cell was observed in SBP group compared to non SBP (P ≤ 0.001, P = 0 β was significantly in-creased in ascitic fluid in patients with SBP versus non SBP (P ≤ 0.001). At cutoff value of 8.3 fl MPV had 85.7% sensitivity and 75% specificity (AUC = 0.876) for diagnosis of SBP. At cutoff value of 15.4 PDW had 90.4% sensitivity and 55% specificity (AUC = 0.762). At cutoff value of 121.9 pg/ml MIP-1β in ascitic fluid had 76.1% sensitivity and 100% specificity (AUC = 0.881) for detecting SBP. Conclusion: MIP-1β and platelet indices are useful marker in the diagnosis of SBP in cirrhotic patients. Combined measurement of MIP-1β in serum and ascitic fluid had 100% sensitivity and specificity for diagnosis of SBP.

Expression of macrophage inflammatory protein-1αin Kupffer cells following liver ischemia or reperfusion injury in rats

瞄准：探索巨噬细胞的表示在在老鼠跟随肝 ischemia/reperfusion 损害 IRI 的 Kupffer 房间(KC ) 的煽动性的 protein-1alpha (MIP-1alpha ) 。方法：四十只男 SD 老鼠随机被划分成五个组。在老鼠肝的部分温暖的 ischemia/reperfusion 损害的一个模型被建立。KC 在灌注以后被孤立并且孵化一个小时，六个小时， 12 h，和 24 h。肿瘤坏死因素高山哈(TNF-alpha ) 并且在上层清液的 interleukin-1beta (IL-1beta ) 被 ELISA 测量。在 KC 的 MIP-1alpha 被细胞化学的免疫和 RT-PCR 检测。结果：没有或很少 MIP-1alpha 蛋白质和 mRNA 在控制组的 KC 被表示。它在 IRI 组的表达式在灌注以后有重要增加(P 【 0.05 ) ，它与控制组相反。结论：在 KC 后面的肝 ischemia/reperfusion 损害的 MIP-1alpha 基因的活跃行为被假定是为肝的 ischemia/reperfusion 损害的主要原因之一。

Anti-oxidative and anti-inflammatory effects of Tagetes minuta essential oil in activated macrophages

Objective:To investigate antioxidant and anti-inflammatory effects of Tagetes minuta(T.minuta)essential oil.Methods:In the present study T.minuta essential oil was obtained from leaves of T.minuta via hydro-distillation and then was analyzed by gas chromatography-mass spectrometry.The antioxidant capacity of T.minuta essential oil was examined by measuring reactive oxygen,reactive nitrogen species and hydrogen peroxide scavenging.The anti-inflammatory activity of T.minuta essential oil was determined through measuring NADH oxidase,inducible nitric oxide synthase and TNF-αmRNA expression in lipopolysacharide-stimulated murine macrophages using realtime PCR.Results:Gas chromatography-mass spectrometry analysis indicated that the main components in the T.minuta essential oil were dihydrotagetone(33.86%),E-ocimene(19.92%).tagetone(16.15%),cis-β-ocimene(7.94%),Z-ocimene(5.27%).limonene(3.1%)and epoxyocimene(2.03%).The T.minuta essential oil had the ability to scavenge all reactive oxygen/reactive nitrogen species radicals with IC_(50)12-15μg/mL,which indicated a potent radical scavenging activity.In addition,T.minuta essential oil significantly reduced NADH oxidase,inducible nitric oxide synthaseand TNF-αmRNA expression in the cells at concentrations of 50μg/mL,indicating a capacity of this product to potentially modulate/diminish immune responses.Conclusions:T.minuta essential oil has radical scavenging and anti-inflammatory activities and could potentially be used as a safe effective source of natural anti-oxidants in therapy against oxidative damage and stress associated with some inflammatory conditions.

Escherichia coli-host macrophage interactions in the pathogenesis of inflammatory bowel disease

Multiple studies have demonstrated alterations in the intestinal microbial community(termed the microbiome) in Crohn's disease(CD) and several lines of evidence suggest these changes may have a significant role in disease pathogenesis. In active and quiescent disease, both the faecal and mucosa-associated microbiome are discordant with matched controls with reduced biodiversity, changes in dominant organisms and increased temporal variation described. Mucosaassociated adherent, invasive Escherichia coli(E. coli)(AIEC), pro-inflammatory and resistant to killing by mucosal macrophages, appear to be particularly important. AIEC possess several virulence factors which may confer pathogenic potential in CD. Type-1 pili(FimH) allow adherence to intestinal cells via cell-surface carcinoembryonic antigen-related cell adhesion molecules and possession of long polar fimbrae promotes translocation across the intestinal mucosa via microfold(M)-cells of the follicle-associated epithelium. Resistance to stress genes(htrA, dsbA and hfq) and tolerance of an acidic pH may contribute to survival within the phagolysosomal environment. Here we review the current understanding of the role of mucosa-associated E. coli in Crohn's pathogenesis, the role of the innate immune system, factors which may contribute to prolonged bacterial survival and therapeutic strategies to target intracellular E. coli.

Macrophage Inflammatory Protein-lalpha mediates Matrix Metalloproteinase-9 enhancement in human adherent monocytes fed with malarial pigment

Objective:To investigate the role of macrophage inflammatory protein-1alpha(MIP-1 alpha) in the detrimental enhancement of matrix mnetalloproteinase-9(MMP-9) expression,release and activity induced by phagocytosis of malarial pigment(haemozoin,HZ) in human monocytes. Methods:Human adherent monocytes were unfed/fed with native HZ for 2 h.After 24 hours. MIP-1 alpha production was evaluated by ELISA in cell supernatants.Alternatively.HZunfed /fed monocytes were treated in presence/absence of anti-human MIP-1 alpha blocking antibodies or recombinant human MIP-lalpha for 15 h(RNA studies) or 24 h(protein studies): therefore,MMP-9 mRNA expression was evaluated in cell lysatcs by Real Time RT-PCR,whereas proMMP-9 and active MMP-9 protein release were measured in cell supernatants by Western blotting and gelatin zvmography.Results:Phagocytosis of HZ by human monocytes increased production of MIP-1 alpha.mRNA expression of MMP-9 and protein release of proMMP-9 and active MMP-9.All the HZ-enbancing effects on MMP-9 were abrogated by anti-human MIP- 1 alpha blocking antibodies and mimicked by recombinant human MIP-l alpha.Conclusions: The present work suggests a role for MIP-lalpha in the HZ-dependent enhancement of MMP-9 expression,release and activity observed in human monocytes.higbligbtiug new detrimental effects of HZ-triggered proinflammatory response by phagocytic cells in falciparum malaria.

Expression of Macrophage Inflammatory Protein 1α in the Endothelial Cells Exposed to Diamide

In order to study whether the endothelial cells (ECs) with lipid peroxidation induced by diamide can express and secrete macrophage inflammatory protein 1α (MIP-1α), the expression of MIP-1α protein in the cells was detected by cell enzyme-linked immunosorbent assay (ELISA) and that of MIP-1α mRNA was determined by cell in situ hybridization and nuclease S1 protection assay after the ECs were exposed to different concentrations of diamide for 4 h. The chemotactic activity of MIP-1α was tested by micropore filter method using modified Boyden chambers. Cell ELISA showed that the expression of MIP-1α protein in endothelial cells exposed to 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was 1.9-fold, 2.3-fold and 1.7-fold respectively as much as that in the control cells, which was statistically significant by analysis of variance. In situ hybridization revealed that the mRNA expression of ECs treated with 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was 1 3-fold, 3.0-fold and 1.7-fold as much as that in the control group, which had statistical significance ( F =188.93, P <0.01). The mRNA expression in 5 μmol/L dimide treated ECs, measured by nuclease S1 protection assay, was 3.4-fold as much as that in the control group( t =8 70, P <0 05). Chemotactic response(99.50±4.31 μm) to the culture medium conditioned by 5 μmol/L diamide treated ECs , which was stronger than that(66.47±3.25 μm) conditioned by the ECs ( F =404.31, P <0.05), was significantly decreased ( F =192.25, P <0.05) after adding MIP-1α antibody. It suggests that diamide, a lipid peroxidation inducer, could stimulate ECs to produce high level of MIP-1α, and might play an important role in atherogenesis by promoting the migration of peripheral blood monocytes into arterial intima.

Macrophages play a role in inflammatory transformation of colorectal cancer

Colorectal cancer(CRC)is one of the most common and fatal cancers worldwide,and it is also a typical inflammatory cancer.The function of macrophages is very important in the tissue immune microenvironment during inflammatory and carcinogenic transformation.Here,we evaluated the function and mechanism of macrophages in intestinal physiology and in different pathological stages.Furthermore,the role of macrophages in the immune microenvironment of CRC and the influence of the intestinal population and hypoxic environment on macrophage function are summarized.In addition,in the era of tumor immunotherapy,CRC currently has a limited response rate to immune checkpoint inhibitors,and we summarize potential therapeutic strategies for targeting tumorassociated macrophages.

Serum macrophage inflammatory protein-lαand high-sensitivity C-reactive protein levels in the paitents with obstructive sleep apnea-hypopnea syndrome and hypertension

Objectives To identify the effects of obstructive sleep apnea-hypopnea syndromemacrophage inflammatory protein -1α(MIP-1α) and high-sensitivity c-reactive protein(hs-CRP) levels,and its impact on cardiac structure and function in patients with hypertension.(OSAHS) on serum. Methods We studied 86 middle-aged subjects classified into four groups according to the absence or presence of OSAHS with and without hypertension.(1)OSAHS patients without hypertension(OSAHS group,n=29);(2)OSAHS patients with hypertension(OSAHS +HT group,n=27);(3) non-OSAHS patients with hypertension(HT group,n =27);(4)volunteers without OSAHS and hypertension(Control subjects, n=27).OSAHS patients were divided into mild,moderate and severe degree based on apnea hypopnea index(AHI).All participants underwent polysomnography and echocardiography. Serum MIP-1αand hs-CRP levels were tested by enzyme linked immunosorbent assay(ELISA).Results Body mass index(BMI),neck collar(NC),waist-to-hip ratio(WHR) in OSAHS group and OSAHS +HT group were significantly higher than those in Control group(PP【0.05).Serum MIP- 1αlevels in OSAHS+HT group was significant higher than HT groups(P【0.05).Serum MlP-1αlevels in those three groups were negative correlationwith AV(r=-0.238,P=0.08) and positively correlated with E/A ratio(r=0.307,P=0.02). Conclusions We have not foundthe cardiac systolic function change in early OSAHS patients with hypertension,while the diastolic function decreased obviously.Serum MIP-1αlevel shows earlier change than hs-CRP level in OSAHS patients which may contribute to the lesion of cardiac diastolic function.

Induction of macrophage inflammatory cytokines by Ox-LDL is ABCA1 dependent

当前的学习试图评估是否的目的由 Ox-LDL 的煽动性的 cytokines 与 ABCA1 的表示有关的巨噬细胞的正式就职小径。巨噬细胞是有处理与 Ox-LDL (30mg/L ) 跟随的 ABCA1 antisense oligonucleotides (100nmol/L ) 的 transfected 的在 THP1/PMA 以后的方法， ABCA1 的表情， ICAM-1 和 MCP-1 mRNA 和蛋白质被决定由即时荧光灯量的 RT-PCR，西方的污点或 ELISA。结果 Ox-LDL 从 THP1/PMA 巨噬细胞在 mRNA 和蛋白质层次导致了 ABCA1， ICAM-1，和 MCP-1 的表情。有 ABCA1 antisense oligonucleotides 的 Transfection 在 12 和 24 个小时以后在 3 和 6 个小时和蛋白质层次以后减少了 ABCA1 mRNA 层次。在由 ABCA1 antisense oligonucleotide 的 ABCA1 蛋白质表示的抑制减少了以后， Ox-LDL 导致的 ICAM-1 和 MCP-1 的表示也被减少。结论巨噬细胞的正式就职由 Ox-LDL 的煽动性的 cytokines 部分依赖于 ABCA1 的表示。我们的研究在巨噬细胞揭示 ABCA1 的新功能。

Toll-like receptor 4-related regulation of Ornithogalum caudatum extract on inflammatory responses in LPS activated macrophages

OBJECTIVE To investigate toll-like receptor 4(TLR4)-related the regulation of Ornithogalum caudatum extract(OCE) on inflammatory responses in lipopolysaccharide(LPS) activated macro.phages.METHODS Primary peritoneal macrophage,Raw 264.7,and THP-1 were incubated in 96-well plate for 24 h and treated with OCE of the concentration of 0-400 μg/ml for 4 h.The viability of cells was measured by MTT assay.Specific concentrations of OCE were added into the medium of primary peri.toneal macrophage,Raw 264.7,and THP-1,respectively,then following with lipopolysaccharides(LPS).Cells were harvested and the total cellular protein and nuclear protein were extracted,and the protein content was determined using BCA protein assay Kit.The expressions of TLR4,inducible nitric oxide synthase(iNOS),cyclooxygenase 2(COX-2),α-inhibitor of NF-κB(IκB-α) and nuclear factor-κB(NF-κB) were assayed by Western blot.The expressions of interleukin-1α(IL-1α),interleukin-1β(IL-1β),interleukin-18(IL-18),and tumor necrosis factor-α(TNF-α) were measured by RT-PCR.RESULTS The results of MTT showed that OCE has no cytotoxicity in Raw 264.7 cells between 1.56 μg/ml and 400 μg/ml.Compared with normal group,the expressions of TLR4,iNOS,COX-2,NF-κB and IL-1α,IL-1β,IL-18,TNF-α,the level of nitric oxide(NO) were significantly increased by LPS stimulation,while OCE pretreat.ment reduced these increase induced by LPS.However,OCE pretreatment reversed the reduction of IκB-α after LPS stimulation.CONCLUSION OCE might suppress TLR4 expression and block the inflamma.tion process of NF-κB and iNOS,further decrease the expression of COX-2 and inhibit the release of inflammatory factors.

Size-dependent macrophage-targeting of mannose-modified rosiglitazone liposomes to alleviate inflammatory bowel disease

Inflammatory bowel disease (IBD) is a refractory chronic intestinal inflammatory disease caused by a malfunction of immune system. As the key immune cells in the intestine, macrophages play an important role in maintaining intestinal homeostasis and tissue repair of the IBD. Pharmacological modulation of macrophage function exhibits the promising therapeutic effect for IBD. In this study, mannose-modified liposomes (MAN-LPs) are prepared for macrophage targeting to improve therapeutic efficiency. Rosiglitazone (ROSI) as an agonist of peroxisome proliferators-activated receptor γ (PPAR-γ) is used as the model drug to fabricate different sized liposomes. The impacts of mannose modification and particle size for macrophage targeting are investigated in cells, zebrafish, and mouse models and the therapeutic effects of the MAN-LPs are evaluated on dextran sulfate sodium (DSS)-induced IBD mouse. Compared to unmodified liposome, MAN-LPs display higher uptake by RAW 264.7 cells and better co-localization with macrophage in zebrafish model. Furthermore, MAN-LPs could effectively accumulate in the inflammatory intestinal sites in IBD mouse model. Most importantly, the targeting ability of MAN-LPs is obviously enhanced with the increasing of particle size, whereas the largest MAN-LPs particles achieve the best anti-inflammatory effect in cells, and a higher therapeutic efficiency in IBD mouse model. Therefore, mannose-modified liposome is a promising strategy for macrophage-targeting in IBD treatment. Particle size of MAN-LPs will affect macrophage targeting ability, as well as the therapeutic effect in-vivo.

Role of macrophages in the progression of acute pancreatitis

In addition to pancreatic cells,other inflammatory cell populations contribute to the generation of inflammatory mediators during acute pancreatitis.In particular,macrophages could be activated by mediators released during pancreatitis by a damaged pancreas.It has been reported that peritoneal macrophages,alveolar macrophages and Kupffer cells become activated in different stages of severe acute pancreatitis.However,macrophages display remarkable plasticity and can change their physiology in response to environmental cues.Depending on their microenvironmental stimulation,macrophages could follow different activation pathways resulting in marked phenotypic heterogeneity.This ability has made these cells interesting therapeutical targets and several approaches have been assayed to modulate the progression of inflammatory response secondary to acute pancreatitis.However,despite the recent advances in the modulation of macrophage function in vivo,the therapeutical applications of these strategies require a better understanding of the regulation of gene expression in these cells.

Functional macrophages and gastrointestinal disorders

Macrophages(MΦ) differe ntiate from blood monocytes and participate in innate and adaptive immunity.Because of their abilities to recognize pathogens and activate bactericidal activities,MΦ are always discovered at the site of immune defense.MΦ in the intestine are unique,such that in the healthy intestine,they possess complex mechanisms to protect the gut from inflammation.In these complex mechanisms,they produce anti-inflammatory cytokines,such as interleukin-10 and transforming growth factor-β,and inhibit the inflammatory pathways mediated by Toll-like receptors.It has been demonstrated that resident MΦ play a crucial role in maintaining intestinal homeostasis,and they can be recognized by their unique markers.Nonetheless,in the inflamed intestine,the function of MΦ will change because of environmental variation,which may be one of the mechanisms of inflammatory bowel disease(IBD).We provide further explanation about these mechanisms in our review.In addition,we review recent discoveries that MΦ may be involved in the development of gastrointestinal tumors.We will highlight the possible therapeutic targets for the management of IBD and gastrointestinal tumors,and we also discuss why more details are needed to fully understand all other effects of intestinal MΦ.